User:Eric M. Walters/Notebook/Spring 2012/2012/06/06

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Checking the 10 mM AA plate for purity

Things were a little sketchy on the 5 mM, which I am tempted to blame on the old primers I'd been going in and out of for a while. Making fresh 1:10 dilutions today and trying colony PCR (internal BC-F w/ outermost acsA-DnRv1) on the 9 patches that came up on the 10 mM acrylate plate plus a gDNA and transformation cassette control.

Each 10 uL reaction:

  • 5 uL GoTaq 2x MM
  • 4 uL Nuclease free water
  • .5 uL fresh BC-F primer
  • .5 uL fresh acsA-DnR primer

This set overnight at 4, and was run Thursday morning.


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